J.G.B. Oostermeijer

Integrating demographic and genetic approaches in plant conservation

Abstract We summarize the problems that populations of formerly common plants may encounter when habitat fragmentation isolates them and reduces population size. Genetic erosion, inbreeding depression, Allee-effects on reproductive success, catastrophes and environmental stochasticity are illustrated with studies on species that have recently become rare in The Netherlands due to habitat fragmentation. These clearly indicate that population viability is negatively affected. We also show that in the recent literature (since 1980), most studies on the conservation of rare plants have addressed population genetic structure and relationships between genetic variation and population size. Though important, these studies are not suitable for assessing the importance of genetics for population viability. In turn, demographic studies can detect changes in vital rates in small populations, but cannot reveal underlying genetic causes. Fitness and demographic studies are also well-represented in the literature, but remarkably few studies have attempted to integrate empirical demographic and genetic studies. We discuss two approaches to fill this very important lacuna in our knowledge. One of these constructs matrix-projection models on the basis of demographic censuses of—if possible—large and viable populations, and combines these with the results of experiments to determine inbreeding effects on demographic transitions and, subsequently, population growth and extinction. The other approach is to demographically monitor experimentally created small and large populations with low and high genetic variation and measure their actual growth rates and probabilities of extinction. We conclude that demography and demographic-genetic experiments should play a central role in plant conservation genetics.

Offspring fitness in relation to population size and genetic variation in the rare perennial plant species Gentiana pneumonanthe (Gentianaceae)

Seeds were sampled from 19 populations of the rare Gentiana pneumonanthe, ranging in size from 5 to more than 50,000 flowering plants. An analysis was made of variation in a number of life-history characters in relation to population size and offspring heterozygosity (based on seven polymorphic isozyme loci). Life-his-tory characters included seed weight, germination rate, proportion of seeds germinating, seedling mortality, seedling weight, adult weight, flower production per plant and proportion of plants flowering per family. Principal component analysis (PCA) reduced the dataset to three main fitness components. The first component was highly correlated with adult weight and flowering performance, the second with germination performance and the third component with seed and seedling weight and seedling mortality. The latter two components were considered as being maternally influenced, since these comprised life-history traits that were significantly correlated with seed weight. Multiple regression analysis showed that variation in the first fitness component was mainly associated with heterozygosity and not with population size, while the third fitness component was only correlated with population size and not with heterozygosity. The latter relationship appeared to be non-linear, which suggests a stronger loss of fitness in the smallest populations. The second (germination) component was neither correlated with population size nor with genetic variation. There was only a weak association between population size, heterozygosity and the population coefficients of variation for each life history character. Most correlation coefficients were negative, however, which suggests that there is more variation among progeny from smaller populations. We conclude that progeny from small populations of Gentiana pneumonanthe show reduced fitness and may be phenotypically more variable. One of the possible causes of the loss of fitness is a combination of unfavourable environmental circumstances for maternal plants in small populations and increased inbreeding. The higher phenotypic variation in small populations may also be a result of inbreeding, which can lead to deviation of individuals from the average phenotype through a loss of developmental stability.

Temporal and Spatial Variation in the Demography of Gentiana Pneumonanthe, a Rare Perennial Herb

1 A total of 35 transition and elasticity matrices for the rare iteroparous herb Gentiana pneumonanthe was analysed for temporal and spatial variation. The data used were collected annually from 10 sites in six different populations on up to seven occasions in the period 1987-93. 2 In general, temporal variation was higher in transition matrices than in elasticities, while between-site variation was high for both transition and elasticity matrices. 3 The relative contributions of three life-history transitions, progression (or growth, G), recruitment from seed (fecundity, F) and survival (retrogression plus stasis, L) to the finite rate of increase, A, were also highly variable, between years within as well as between sites. The observed variation is very large in comparison with that previously observed either between or within other iteroparous herbs from open habitats, even showing some overlap with demographic patterns normally characteristic of woody plants. 4 A G-L-F ordination shows a long, narrow band across the entire diagram from matrices with a low elasticity for L and high elasticities for G and F at one end to matrices which have an elasticity of 1.00 for L on the other. 5 Correlations between the G-, Land F-elasticities, A, and the vegetation cover suggest that the band in the ordination diagram represents a successional pathway in wet heathlands from invasive to regressive populations. Elasticity matrices from regularly mown hay meadows are characteristic of stable populations, but represent senile, regressive populations after mowing has ceased. 6 Relationships between A and the elasticities of G, L and F show that in matrices of stable or declining populations (A 1) the contribution of progression and fecundity becomes larger. 7 Given their relatively large temporal and spatial variation, elasticities are useful in nature conservation and management only if the corresponding value of A is taken into account. 8 Significant positive correlations between transition probabilities of different life stages were observed. This phenomenon may increase the risk of extinction by environmental stochasticity.

Population structure of the rare, long-lived perennial Gentiana pneumonanthe in relation to vegetation and management in The Netherlands

After analysis of population age-state spectra of Gentiana pneumonanthe in a variety of vegetation types, three different population types could be distinguished: (a) «invasive» or «dynamic» populations, characterized by high densities of seedlings and juveniles relative to the adult age states, (b) «normal» or «stable» populations with adult age states prevailing, but with low densities of seedlings and juveniles, and (c) «regressive» or «senile» populations, consisting only of adult flowering and adult vegetative individuals. The structure of the surrounding vegetation appears to be very important in determining the population structure of the marsh gentian. In particular the percentage of bare soil surface and the cover of the litter layer explain most of the variation in the density and proportion of different age states in the populations

Analysis of the relationship between allozyme heterozygosity and fitness in the rare Gentiana pneumonanthe L.

Especially for rare species occurring in small populations, which are prone to loss of genetic variation and inbreeding, detailed knowledge of the relationship between heterozygosity and fitness is generally lacking. After reporting on allozyme variation and fitness in relation to population size in the rare plant Gentiana pneumonanthe, we present a more detailed analysis of the association between heterozygosity and individual fitness. The aim of this study was to test whether increased fitness of more heterozygous individuals is explained best by the ‘inbreeding’ hypothesis or by the ‘overdominance’ hypothesis. Individual fitness was measured during 8 months of growth in the greenhouse as the performance for six life‐history parameters. PCA reduced these parameters to four main Fitness Components. Individual heterozygosity was scored for seven polymorphic allozyme loci.

Myrmecochory in Polygala vulgaris L., Luzula campestris (L.) DC. and Viola curtisii Forster in a Dutch dune area

SummarySeed dispersal by ants in Polygala vulgaris, Luzula campestris and Viola curtisii was studied in a primary dune valley on the island of Terschelling, The Netherlands. Normally developed seeds of all three species are taken by the ants into their nests. The ants show a distinct preference for the seeds of the specialized myrmecochore Polygala vulgaris, as compared with the two diplochorous species. It could be demonstrated that the elaiosome is the attractive part of the seed. Mapping studies demonstrate that the dispersal of the seeds by ants has a marked effect on the distribution pattern of the standing population of Polygala and Viola. Adult plants are often found on or close to the active nest mounds of all ant species present, while the growing sites of juvenile individuals and seedlings are practically restricted to the nest environment. The nests of two of the seed-dispersing ant species, viz., those of Lasius niger and Tetramorium caespitum, show differences in soil chemistry with the surroundings. The ant nests are significantly richer in some essential plant macronutrients, such as phosphate, potassium and nitrate. The advantage of myrmecochory in the dune area of Terschelling is discussed.

Reproductive success and clonal genetic structure of the rate Arnica montana (Compositae) in The Netherlands

In a medium-sized population ofArnica montana, a threatened species in The Netherlands, the breeding system, reproductive success and genetic clonal structure were studied. Pollination experiments suggested thatA. montana is largely self-incompatible. Inbreeding depression was observed for seedling weight but not for fruit weight and germination rate. Although genetic variation is rather low in this population, the data suggest an outcrossing mating system. Analysis of the genotype of all mapped rosettes in a plot of 100 m2 indicated that dense clusters often consist of identical genotypes, suggesting a clonal structure. Open clusters frequently contained several different genotypes. This may be caused by limited fruit dispersal, since seedlings were found mainly within or in the near surroundings of the clusters.

Dispersal limitation in epiphytic bromeliad communities in a Costa Rican fragmented montane landscape

Transformation of tropical forests is likely to affect seed-dispersal patterns and influence the composition of epiphytic plant communities in human-altered habitats. We tested this hypothesis by carrying out a comparative study of seed influx, survival and growth of transplanted seedlings of epiphytic bromeliads among isolated trees in six pasture areas, six forest edges and six forest sites in a montane area in Costa Rica. In total, 72 traps trapped 1285 seeds over a 2-mo period in the dry season of 2003. For all four investigated bromeliad genera, Catopsis, Guzmania, Tillandsia and Werauhia, the number of trapped seeds in each habitat followed a pattern similar to the numberoffruitingindividualsinthevicinityofthetraps.Trapsinforestedges(30)were1.9timesmorelikelytocollect seeds than traps at forest interiors (30) and pasture trees (12), the latter showing similar probabilities of catching seeds. After 1 y, survival and growth of 3660 transplanted seedlings from three bromeliad species was significantly higher in forest interiors, providing no explanation for the lower abundance of fruiting adults in that habitat. These resultssuggestthatthesuccessfulestablishmentofepiphyticbromeliadsinforestinteriorsismainlydispersal-limited.If corroborated,differencesinabundanceamongspeciesateachhabitatarelikelyrelatedtodifferencesingrowthratesand reproductive success. Further studies on the growth and mortality of seedlings up to theflowering stage, however, are needed.

Future prospects for the rare, late-flowering Gentianella germanica and Gentianopsis ciliata in Dutch nutrient-poor calcareous grasslands

We discuss the population biology of two calcareous grassland gentians, Gentianella germanica and Gentianopsis ciliata, in relation to the habitat management currently practiced in The Netherlands. There, at the margin of their range, both species are rare. Gentianella germanica persists on six remaining locations, whereas Gentianopsis ciliata, with two populations, is nearly extinct. Gentianella germanica is a strict biennial, Gentianopsis ciliata an iteroparous perennial. Both species depend on insects for seed production and su!er from low insect visitation. Pollination experiments in one Dutch population demonstrated that Gentianopsis ciliata is self-compatible, but hardly sets any seed under natural conditions due to pollen limitation. The low reproductive success of both species is partly due to the low pollinator densities at their late flowering time, partly caused by the small population size of the gentians themselves, and partly a result of mowing too early. The latter has destroyed the seed crop of several subsequent years in one population of Gentianopsis ciliata and some of Gentianella germanica. In Gentianella germanica, the early mowing and low insect visitation seems to have resulted in selection of less herkogamous and consequently more autofertile individuals. The perspectives for Gentianopsis ciliata are currently extremely poor in The Netherlands. Under the present circumstances, extinction will most likely occur within 10‐20 years. Population reinforcement (seeding, artificial cross-pollination with nearby populations) should be considered if we want to conserve this species. For both gentians, but also of other flagship species of nutrient-poor calcareous grasslands, the total grassland area needs to be enlarged and must constitute an interconnected network of reserves. The traditional management method, sheep grazing, is to be preferred over mowing, but only if the flock visits each grassland patch or reserve for only a short time of less than a day, and with intervals of more than 2 weeks between visits. If mowing is preferred for other reasons, it should be done rotationally, and not before October. # 2002 Elsevier Science Ltd. All rights reserved.

Existence of vigorous lineages of crop-wild hybrids in Lettuce under field conditions.

Plant to plant gene flow is a route of environmental exposure for GM plants specifically since crosses with wild relatives could lead to the formation of more vigorous hybrids, which could increase the rate of introgression and the environmental impact. Here, we test the first step in the process of potential transgene introgression: whether hybrid vigor can be inherited to the next generation, which could lead to fixation of altered, i.e., elevated, quantitative traits. The potential for a permanent elevated fitness was tested using individual autogamous progeny lineages of hybrids between the crop Lactuca sativa (Lettuce) and the wild species Lactuca serriola (Prickly Lettuce). We compared progeny from motherplants grown under either greenhouse or field conditions. The survival of young plants depended strongly on maternal environment. Furthermore, we observed that offspring reproductive fitness components were correlated with maternal fitness. Our study demonstrates that post-zygotic genotypic sorting at the young plants stage reduces the number of genotypes non-randomly, leading to inheritance of high levels of reproductive traits in the surviving hybrid lineages, compared to the pure wild relatives. Consequently, directional selection could lead to displacement of the pure wild relative and fixation of more vigorous genome segments originating from crops, stabilizing plant traits at elevated levels. Such information can be used to indentify segments which are less likely to introgress into wild relative populations as a target for transgene insertion.